清热透表方对流感病毒小鼠TLR7、MyD88、NF-κB、AP-1 mRNA及蛋白的表达

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目的观察解表清肺方对甲型流感病毒FM1感染小鼠肺组织细胞Toll样受体7(Toll-like receptor 7,TLR7)髓样分化因子88(myeloid differentiation factor 88,My D88)、激活转录因子核因子-Kappa B(nuclear factor-kappa B,NF-κB)、转录激活因子-1(transcription activator-1,AP-1)mRNA及蛋白表达影响。方法将C57小鼠分为6组,每组8只,将其随机分为正常组,模型组,阳性对照组(即利巴韦林组,剂量为0.5g·kg-1·d-1),解表清肺方高、中、低剂量组(即中药高、中、低剂量组,其剂量分别为8,4,2g·kg-1·d-1),将各组小鼠乙醚轻度麻醉,滴鼻感染15LD50流感病毒液,每鼠4滴,约0.05 ml,正常组以生理盐水滴鼻,于攻毒前一天开始灌胃给药,共给药5 d,每天1次,每次0.3 ml,正常组和模型组分别给予等溶剂的蒸馏水,于攻毒第4天处死小鼠。采用RT-PCR反应测定小鼠肺组织的TLR7、My D88、NF-κB、AP-1 mRNA的表达,采用western blot法测定测定肺组织TLR7、My D88、NF-κB、AP-1蛋白表达水平。结果与正常组相比,模型组TLR7、My D88、NF-κB、AP-1 mRNA及蛋白表达升高(P<0.01),阳性对照组,中药各剂量组与模型组相比TLR7、My D88、NF-κB、AP-1 mRNA表达均有降低(P<0.01或P<0.05),而中药高中剂量组TLR7、My D88、NF-κB、AP-1的蛋白表达较模型组相比有降低(P<0.01或P<0.05),而中药低剂量组与模型组相比TLR7,NF-κB的蛋白表达有降低,但My D88、AP-1却无显著差异(P>0.05)。结论清热透表方各剂量组可以通过以My D88依赖的TLR7为主的信号通路,发挥抗流感病毒的作用。 Objective To observe the effect of JDPDP on myeloid differentiation factor 88 (Myeloid differentiation factor 88, TLR7) in lung tissue of mice infected with influenza A virus FM1, (NF-κB), transcriptional activator-1 (AP-1) mRNA and protein expression in liver of rats. Methods C57 mice were divided into 6 groups with 8 mice in each group. They were randomly divided into normal group, model group and positive control group (ribavirin 0.5g · kg-1 · d-1) , Respectively. The mice in each group were given high, medium and low doses of Chinese herbal medicine (namely, high, medium and low doses of traditional Chinese medicine at dose of 8,4 and 2 g · kg-1 · d-1, respectively) Degree of anesthesia, intranasal infection of 15LD50 influenza virus solution, 4 drops per mouse, about 0.05 ml, normal saline nasal drops, one day before challenge intragastric administration, a total of 5 d administration, once a day, each Times 0.3ml, the normal group and the model group were given an equal solvent of distilled water, the mice were killed on the fourth day of challenge. The expression of TLR7, My D88, NF-κB and AP-1 mRNA were detected by RT-PCR. The expression of TLR7, My D88, NF-κB and AP- . Results Compared with the normal group, the mRNA and protein expressions of TLR7, My D88, NF-κB and AP-1 in model group were significantly increased (P <0.01). Compared with model group, TLR7, My D88 (P <0.01 or P <0.05), while the protein expression of TLR7, My D88, NF-κB and AP-1 in high school and middle dose group of TCM decreased compared with the model group (P <0.01 or P <0.05). Compared with the model group, the expression of TLR7 and NF-κB in the low dose group of traditional Chinese medicine decreased, but there was no significant difference between My D88 and AP-1 (P> 0.05). CONCLUSION: Each dose group of Qingre Tongpi can exert the anti-influenza virus effect through the My D88-dependent TLR7-based signaling pathway.
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